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盐碱胁迫对尼罗罗非鱼鳃Na+/HCO3-共转运子、碳酸酐酶基因表达的影响
引用本文:梁从飞,赵金良,甘远迪,王飞,Thammaratsuntorn Jeerawat,伍勇,李传阳,罗明坤.盐碱胁迫对尼罗罗非鱼鳃Na+/HCO3-共转运子、碳酸酐酶基因表达的影响[J].中国水产科学,2016,23(2):274-283.
作者姓名:梁从飞  赵金良  甘远迪  王飞  Thammaratsuntorn Jeerawat  伍勇  李传阳  罗明坤
作者单位:上海海洋大学农业部淡水水产种质资源重点实验室,上海,201306
基金项目:国家科技支撑计划项目(2012BAD16B03),现代农业产业技术体系专项(CARS-49),水产动物遗传育种中心上海市协同创新中心项目(ZF1206)
摘    要:为了探讨盐碱胁迫条件下鱼类渗透生理调节机制,以尼罗罗非鱼(Oreochromis niloticus)为实验材料, PCR扩增得到了Na+/3HCO-共转运子(NBCe1)基因cDNA部分序列,比较了单盐(盐度10、盐度15)、单碱(1.5 g/L、3 g/L NaHCO3)、盐碱混合(盐度10,碱度1.5 g/L;盐度15,碱度3 g/L)胁迫后不同时间(0 h、6 h、12 h、24 h、48 h、72 h、96 h)血清渗透压、离子浓度(Na+、K+、Cl–、Ca2+)以及鳃碳酸酐酶(CA)活性、CA与NBCe1基因mRNA表达变化。结果显示,不同胁迫条件下,血清渗透压、离子浓度、鳃组织 CA 酶活、CA 与 NBCe1基因 mRNA 表达变化均与胁迫强度呈正相关。随时间推移,血清渗透压、离子浓度呈现先上升后下降的变化趋势,单盐、盐碱混合组血清渗透压值较单碱组高。单盐、单碱、盐碱混合组中, NBCe1基因mRNA在鳃中均呈略微上调,但不显著(P>0.05)。单碱组和盐碱混合组鳃CA活性较单盐组高,低盐碱胁迫(盐度10,碱度1.5 g/L)下CA活性较晚达最高值;不同胁迫条件下, CA基因mRNA表达均表现上调,单碱、盐碱混合组更为显著(P<0.05),推测CA较NBCe1对体内3HCO-转运作用更为显著。研究结果为尼罗罗非鱼盐碱适应生理调节提供了基础资料。

关 键 词:尼罗罗非鱼  盐碱胁迫  Na%2B/3HCO?共转运子  碳酸酐酶  酶活性  mRNA表达

Effects of salinity and alkalinity on mRNA expression of Na+/HCO3- cotransporter and carbonic anhydrase genes from Oreochromis niloticus
LIANG Congfei,ZHAO Jinliang,GAN Yuandi,WANG Fei,Thammaratsuntorn Jeerawat,WU Yong,LI Chuanyang,LUO Mingkun.Effects of salinity and alkalinity on mRNA expression of Na+/HCO3- cotransporter and carbonic anhydrase genes from Oreochromis niloticus[J].Journal of Fishery Sciences of China,2016,23(2):274-283.
Authors:LIANG Congfei  ZHAO Jinliang  GAN Yuandi  WANG Fei  Thammaratsuntorn Jeerawat  WU Yong  LI Chuanyang  LUO Mingkun
Institution:Laboratory of Freshwater Fisheries Germplasm Resource, Ministry of Agriculture;Shanghai Ocean University, Shanghai 201306, China
Abstract:To understand fish osmotic adjustment mechanisms in saline and alkaline water, the partial cDNA se-quence was obtained from gills ofOreochromis niloticus. Physiological changes in serum osmolality, ion concen-tration (Na+, K+, Cl– and Ca2+), and gill carbonic anhydrase (CA) activities were determined, andCAandNBCe1 mRNA gene expressions under saline (10 g/L, 15 g/L NaCl), alkaline (1.5 g/L and 3 g/L NaHCO3), and sa-line-alkaline (salinity 10, 15 g/L NaCl; salinity 1.5, 3 g/L NaHCO3) conditions at different times (0 h, 6 h, 12 h, 24 h, 48 h, 72 h and 96 h) were compared. The results showed that serum osmolality, ion concentration, gill CA activity, CA andNBCe1 mRNA gene expression correlated positively with the strength of saline, alkaline and sa-line-alkaline stress. Over time, serum osmolality and ion concentration trends increased and then decreased. Os-motic pressure insaline and saline-alkaline water was higher than that in alkaline water. Gill CA activity in alkaline and saline-alkaline water was higher than that in saline water. Under low concentrations of stressors, CA activity reached its highest level at a later time. Slightly higherNBCe1 gene mRNA expression was detected in gills under high concentrations of stressors (P>0.05). GillCA mRNA expression in saline, alkaline and saline-alkaline water was increased, but the increase was more evident in alkaline and saline-alkaline water (P<0.05). The results showed that CA and NBCe1 in Nile tilapia are involved in salinity and alkalinity regulation under osmotic stress. The results provide a basic understanding of the physiological regulation during salinity-alkalinity adaptation.
Keywords:Oreochromis niloticus  salt and alkline stress  Na+/3HCO? cotransporter  carbonic anhydrase  enzyme activity  mRNA expression
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